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Effect of maternal micronutrients (folic acid and vitamin B12) and omega 3
fatty acids on indices of brain oxidative stress in the offspring

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Original article

Maternal micronutrients (folic acid and vitamin B12) and omega

3 fatty acids: Implications for neurodevelopmental
risk in the rat offspring
Suchitra Roy 1, Anvita Kale 1, Kamini Dangat, Pratiksha Sable,
Asmita Kulkarni, Sadhana Joshi ⇑
Interactive Research School for Health Affairs, Bharati Vidyapeeth University, Pune 411043, India

Received 15 September 2010; received in revised form 5 January 2011; accepted 8 January 2011

Abstract

Altered maternal micronutrients (folic acid, vitamin B12) are suggested to be at the heart of intra-uterine programming of adult
diseases. We have recently described interactions of folic acid, vitamin B12 and docosahexaenoic acid in one carbon metabolism that
is considered to play a key role in regulation oxidative stress and chromatin methylation. However its impact on fetal oxidative stress
and brain fatty acid levels has been relatively unexplored. The present study examined the effect of imbalance in maternal micro-
nutrients (folic acid and vitamin B12) and maternal omega 3 fatty acid supplementation on oxidative stress parameters and brain
fatty acids and in the offspring at birth. Pregnant female rats were divided into six groups at two levels of folic acid both in the
presence and absence of vitamin B12. Both the vitamin B12 deficient groups were supplemented with omega 3 fatty acid. Oxidative
stress marker (malondialdehyde) and polyunsaturated fatty acid profiles in plasma and brain were analyzed in dam and offspring at
d20. Our results for the first time indicate that imbalance in maternal micronutrients (excess maternal folic acid supplementation on
a B12 deficient diet) increases (p < 0.01) oxidative stress in both mother and pups. This increased maternal oxidative stress resulted in
lower (p < 0.01) fetal brain DHA levels. Omega 3 fatty acid supplementation was able to restore (p < 0.05) the levels of brain DHA
in both the vitamin B12 deficient groups. Our data has implications for implications for neurodevelopmental disorders since micro-
nutrients and DHA are important modulators for neural functioning.
Ó 2011 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

Keywords: Docosahexaenoic acid; Folic acid; Micronutrients; Oxidative stress; Vitamin B12

1. Introduction developmental origins of disease has been on the avail-

It is well established that the environment encoun-
tered during fetal life and infancy is strongly related
to increased risk of non-communicable diseases in
adult life [1]. Although much of the focus on the
⇑ Corresponding author. Address: Interactive Research School for
Health Sciences, Bharati Vidyapeeth University, Pune Satara Road,
Pune 411043, India. Tel.: +91 020 24366929x31; fax: +91 020 24366929.
E-mail address: srjoshi62@gmail.com (S. Joshi).
1
These authors contributed equally to this work.
ability of energy and protein during these critical devel-
opmental periods, micronutrient deficiencies may also
play an important role in fetal growth and development
[2]. In developing countries, like India micronutrient
deficiencies are common and associated with poor
pregnancy outcomes [3]. Importance of maternal folic
acid in prevention of neural tube defects is well estab-
lished [4]. In this view, in India, there is a policy for
supplementing all pregnant women with iron and folic
acid (60 mg and 500 lg per day) for prevention of neural
tube defects [5].

0387-7604/$ - see front matter Ó 2011 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.braindev.2011.01.002
 S. Roy et al. / Brain & Development 34 (2012) 64–71
Many countries like USA, Canada and Chile have
also introduced folic acid fortification program wherein
all flour and uncooked cereal-grain products were forti-
fied with folic acid [6–9]. This folic acid fortification has
been shown to improve the folate status and reduce the
prevalence and incidence of neural tube defects [10–15].
However, the long-term effect of folic acid fortification
on the risk of colorectal cancer and other malignancies
remain unclear [16]. Recent studies have raised concerns
that folic acid fortification may mask symptoms as well
as worsen the vitamin B12 deficiency and lead to adverse
neurologic consequences [17,18]. Most of the Indian
women during pregnancy are known to have lower vita-
min B12 levels [19] and is therefore a cause of concern.
Our own studies in animals and by others have shown
that folic acid alters levels of omega 3 fatty acids [20]. A
series of our studies in pregnancy have well established
the importance of omega 3 fatty acids especially DHA
during pregnancy [21–25]. Docosahexaenoic acid
(DHA), an omega 3 polyunsaturated fatty acid highly
enriched in the brain, is vital for proper brain develop-
ment and function [26]. Micronutrient insufficiency or
imbalance during pregnancy can compromise brain
development and function. Our studies in young schizo-
phrenic patients have also shown altered micronutrients
and DHA at the onset of psychosis implicating a neuro-
developmental pathophysiology in schizophrenia [27].
This has led us recently to describe that in one carbon
metabolic pathway, membrane phospholipids are major
methyl group acceptors and reduced DHA levels may
result in diversion of methyl groups towards DNA ulti-
mately resulting in DNA methylation [28]. Reports sug-
gest that many neurological disorders like childhood
cognitive dysfunction, schizophrenia which become
apparent after birth have their origin during fetal life
[29–32].
We hypothesize that altered maternal intake and
metabolism of folic acid, vitamin B12 and omega 3 fatty
acids in one carbon metabolism and induction of oxida-
tive stress during pregnancy may lead to reduced levels
of brain LCPUFA in the offspring at birth which may
have implications for neurodevelopmental disorders in
later life. The present study for the first time examines
the effect of (1) imbalance of maternal micronutrients
(folic acid and vitamin B12) and (2) maternal omega 3
fatty acid supplementation in the above diets on brain
fatty acids and oxidative stress parameters in the off-
spring at birth.
2. Materials and methods
All experimental procedures were in accordance with
guidelines of Institutional Animal Ethics Committee
(IAEC). The institute is recognized to undertake exper-
iments on animals as per the committee for the purpose
65
of control and supervision of experimental animals,
Govt of India (No. 258/CPCSEA).
2.1. Animals
Wistar albino rats (60F, 15M) of average weight 150 g
were obtained from National Toxicology Center animal
house. Instead of using them directly for the experimental
protocol it was thought appropriate to use their progeny.
They were maintained at 22 °C on a controlled 12-h light
and 12 h dark cycle with appropriate ventilation system.
Animals were marked with picric acid as Head (H), Back
(B), Tail (T), etc. for identification.
2.2. Breeding
These pups were then put for breeding at 3 months of
age. Males were housed individually prior to mating to
acquire cage dominance. Virgin female rats were
allowed to breed (sex ratio 1:3). On the following morn-
ing the vaginal smears were taken to confirm mating.
Vaginal smears were taken on a clean microscope slide
using a cotton bud dipped in saline. The slides were
examined under a microscope at 10 magnification.
The sperm positive smear was considered a result of suc-
cessful mating and considered day 0 of gestation. The
pregnant dams were housed individually (in polypropyl-
ene cages of 29  22  14 cm dimensions containing rice
husk as bedding material) and allowed to deliver nor-
mally. Animals receiving Vitamin B12 deficient diets
were kept in special cages to prevent coprophagy.
Out of 60 females, 47 females became pregnant and
divided randomly into six dietary groups. All dams were
delivered by C section on d20 of gestation. Two pups
from each group were randomly selected and analyzed
for organ weights and biochemical parameters.
2.3. Diets
The composition of the control and the treatment
diets was as per AIN 93 purified diets for laboratory
rodents [33]. Protein level in the control and treatment
diets was 18%. Total of six isocaloric treatment diets
were formulated. Four diets were formulated for exam-
ining the effects of two (i.e. 2 and 8 mg folic acid/kg diet)
different levels of folic acid during pregnancy both in the
presence and absence of vitamin B12. In addition, two
more diets were formulated to examine the effects of
omega 3 fatty acid (DHA + EPA) supplementation on
both the vitamin B12 deficient groups. Vitamin B12 defi-
ciency was obtained exclusively through dietary means.
The diets contained the following ingredients (g/kg
diet) (casein, 200; corn starch, 398; dextrinized starch,
132; sucrose, 100; fiber, 50; mineral mixture, 35; vitamin
mixture, 10; cystine, 3; choline bitartarate, 2.5; tertiary
butyl hydroxyl quinone, 0.014; soyabean oil, 70).
66 S. Roy et al. / Brain & Development 34 (2012) 64–71
Vitamin free casein was used for all treatment diets. The
mineral mixture composed of the following ingredients
(g/kg mixture): calcium carbonate, 357; potassium phos-
phate, 196; potassium citrate, 70.78; sodium chloride,
78; potassium sulphate, 46.6; magnesium oxide, 24; fer-
ric citrate, 6.06; zinc carbonate, 1.65; manganous car-
bonate, 0.63; cupric carbonate, 0.3; potassium iodate,
0.01; sodium selenate, 0.01; ammonium paramolybdate,
0.007; sodium metasilicate, 1.45; chromium potassium
sulphate, 0.275; lithium chloride, 0.01; boric acid, 0.08;
sodium fluoride, 0.06; nickel carbonate, 0.03; ammo-
nium vanadate, 0.006 and sucrose, 221.02. The vitamin
mixture composed of the following ingredients (g/kg
mixture): nicotinic acid, 3; calcium pantothenate, 1.6;
pyridoxine–HCl, 0.7; thiamin–HCl, 0.6; riboflavin, 0.6;
D-Biotin, 0.02; vitamin B12 (in 0.1% Mannitol), 2.5; vita-
min E, 15; vitamin A, 0.8; vitamin D-3, 0.25; vitamin K,
0.075; folic acid, 0.2 (control) and sucrose 974.655, was
used to make total weight of the vitamin mixture to
1 kg.
Thus there were a total of six groups, control
(NLFolate + NLB12 i.e. normal folate, normal B12);
NLFolate + noB12 (normal folate, B12 deficient); NLFo-
late + noB12 + omega 3 supp (normal folate, B12 defi-
cient, omega 3 supplemented); HFolate + NLB12
(High folate, normal B12); HFolate + noB12 (High
folate, B12 deficient) and HFolate + noB12 + omega 3
supp (High folate, B12 deficient, omega 3 supplemented).
The lowest level i.e. 2 mg/kg represents the normal
level of folic acid used in the control diet as per the cur-
rent AIN 93 guidelines while 8 mg/kg is roughly four
times the requirement of a normal rat. This is in accor-
dance with the fact that folic acid requirement for
Indian pregnant woman is set at 400 lg/d which is four
times the requirement of a non-pregnant woman. The
level of omega 3 fatty acid supplementation was chosen
to have an omega 6/omega 3 ratio of 1:1 which is consid-
ered to be the ideal ratio [34].
All diets contained tertiary butyl hydroquinone
(TBHQ) to prevent oxidation [33,35,36]. Diets were
stored at refrigeration temperatures and fresh diet was
provided daily.
2.4. Observations recorded
During pregnancy, dam weights were recorded at 0,
7, 14 and 20 d to obtain weight gains. On d20 of gesta-
tion the litter weight and size was recorded in each
group.
2.5. Brain weights
The pups were cut open from the gestational sac and
weighed before dissecting. Then the pups were dissected
to collect the whole brain and the absolute weight of the
brain was recorded on a Schimadzu electronic balance
with a least count of 0.001 g and maximum capacity of
220 g. These brains were immediately snap frozen in
liquid nitrogen and stored at 80 °C for biochemical
estimations. The relative brain weights were expressed
in relation to the pup weights i.e. (absolute brain
weight/pup weight  100).
2.6. Analysis of fatty acids
Whole brains of the pups were homogenized in cold
phosphate buffer saline, pH 7.5 using a Teflon glass
homogenizer on ice. The homogenate was centrifuged at
10,000 rpm at 4 °C for 20 min. The membranes were
diluted in 2 ml phosphate buffer saline and stored at
20 °C. Fatty acids were analyzed as methyl esters by
the method as recently described by us [37]. Briefly,
transesterification of the phospholipid fraction was
carried out using HCl–methanol. Methyl esters were
separated and quantified using a Perkin-Elmer gas chro-
matograph (SD 2330, 30 m capillary column, Supelco,
PA, USA). Helium was used as carrier gas at 1 ml/min.
Oven temperature was held at 150 °C for 10 min pro-
grammed to rise from 150 to 220 °C at 10 °C/min and at
220 °C for 10 min. The detector temperature was 275 °C
and the injector temperature was 240 °C. Retention times
and peak areas were automatically computed. Individual
fatty acids were identified by comparison of peaks with
peaks of standard fatty acid methyl esters (Sigma,
USA). The data included is on only critically important
long chain polyunsaturated fatty acids and major fatty
acids. The omega 3 fatty acids included alpha linolenic
acid, eicosapentaenoic acid and docosahexaenoic acid
while omega-6 fatty acids included linolenic acid, gamma
linolenic acid, dihomogammalinolenic acid, docosapen-
taenoic acid and arachidonic acid. The monounsaturated
fatty acids included myristoleic acid, palmitoleic acid,
oleic acid and nervonic acid. Saturated fatty acids
included myristic acid, palmitic acid and stearic acid.
2.7. Lipid peroxidation measurements (dam plasma and
pup brain MDA levels)
Malondialdehyde (MDA) levels were estimated from
dam plasma and pups brain using kit from Bioxytech
MDA-586 kit (Oxis International, USA). Briefly, thio-
barbituric acid reacts with malondialdehyde to form
pink color, and the absorbance was measured at
586 nm. Tetramethoxypropane is used as a standard.
Brain MDA concentration is expressed as nmol/ml.
2.8. Statistical methods
Litter means were used as the unit of analysis. Values
are mean ± SD. The data were analyzed using SPSS/
PC + package (Version 11.0, Chicago, IL). Mean values
of the estimates of various parameters for the treatment
 S. Roy et al. / Brain & Development 34 (2012) 64–71
groups were compared with those of control group at
conventional levels of significance, using least signifi-
cance difference estimated from one way analysis of var-
iance (ANOVA).
3. Results
3.1. Feed intake
The daily feed intake was (19.44 ± 1.86 g) for control;
(17.80 ± 3.12 g) for NLFolate + noB12; (18.22 ± 2.14 g)
for HFolate + NLB12 and (18.49 ± 3.72 g) for HFo-
late + noB12 groups suggesting that imbalance in maternal
micronutrients (folic acid and vitamin B12) did not affect
feed intake. In contrast, feed intake in both the omega 3
fatty acid supplemented groups was lower (p < 0.01) i.e.
NLFolate + noB12 + omega 3 supp (14.91 ± 1.33 g) and
HFolate + noB12 + omega 3 supp (15.32 ± 2.15 g) than
control. Further feed intake in omega 3 fatty acid supple-
mented groups were also lower than their respective B12
deficient groups (NLFolate + noB12 vs. NLFolate +noB
12 + omega 3 supp, p < 0.05) and (HFolate + noB12 vs.
HFolate + noB12 + omega 3 supp, p < 0.05).
3.2. Reproductive performance
Weight gain of dams during pregnancy was similar in
all the groups i.e. control (106.5 ± 19.97), NLFolate +
noB12 (111.75 ± 20.10), HFolate + NLB12 (103.00 ±
37.78) and HFolate + noB12 (113.50 ± 26.30). Further
supplementation of omega 3 fatty acids to the vitamin
B12 deficient groups was also similar i.e. NLFolate +
noB12 + omega 3 supp (107.87 ± 13.88) and HFolate +
noB12 + omega 3 supp (103.75 ± 19.36). The pup weight
was comparable between all treatment groups i.e.
control (3.51 ± 0.92), NLFolate + noB12 (3.74 ± 0.61),
HFolate + NLB12 (3.61 ± 1.23), HFolate + noB12
(3.36 ± 0.51), NLFolate + noB12 + omega 3 supp (3.15 ±
0.81) and HFolate + noB12 + omega 3 supp (3.40 ±0.41).
3.3. Relative brain weights
There was no difference in relative brain weights
of pups among all the groups i.e. control (0.04 ±
0.007), NLFolate + noB12 (0.04 ± 0.005), HFolate +
NLB12 (0.04 ± 0.008), HFolate + noB12 (0.04 ± 0.008),
NLFolate + noB12 + omega 3 supp (0.04 ± 0.01) and
HFolate + noB12 + omega 3 supp (0.04 ± 0.001).
3.4. Dam plasma and pup brain MDA levels
Dams plasma MDA levels from the HFolate + noB12
group was higher (p < 0.01) as compared to control,
NLFolate + noB12 and HFolate + NLB12 groups
(Fig. 1). However, plasma MDA levels in dams from the
other groups was comparable to control. Pup brain
67
MDA from the HFolate + noB12 group was also higher
(p < 0.01) as compared to control and HFolate + NLB12
groups (Fig. 1).
3.5. Dam plasma fatty acids
DHA and AA were comparable in the HLFolate +
NLB12 group as compared to control. However, DHA
levels in the HLFolate + noB12 group was lower
(p < 0.05) as compared to control (Fig. 1). Supplementa-
tion of omega 3 fatty acids to the NLFolate + noB12 and
HLFolate + noB12 groups improved levels of DHA and
reduced levels of AA (p < 0.01 for both) as compared to
control. Further, DHA levels in HLFolate + noB12 +
omega 3 supp was higher and AA levels were lower as
compared to HLFolate + noB12 group.
3.6. Pup brain fatty acids
In the pup brain DHA levels were lower in both the
NLFolate + noB12 and HLFolate + noB12 groups (p <
0.01 for both) as compared to control (Fig. 1).
Reductions in DHA (p < 0.05) were also seen in the
HLFolate + NLB12 group as compared to control. In
contrast, both NLFolate + noB12 + omega supp and
HLFolate + noB12 + omega supp groups showed
improved DHA levels (p < 0.01 for both). However, ara-
chidonic acid levels were reduced (p < 0.05) as compared
to control and their respective B12 deficient groups.
4. Discussion
This is the first report which has examined the effect of
imbalance of maternal micronutrients (folic acid and
vitamin B12) and omega 3 fatty acid supplementation
on maternal oxidative stress marker (malondialdehyde)
and pup brain LCPUFA levels on d20 of gestation.
Our results indicate (1) imbalance in maternal micronu-
trients like folic acid and vitamin B12 or omega 3 fatty
acid supplementation did not affect weight gain during
pregnancy. (2) Maternal folic acid supplementation in
the presence of vitamin B12 did not alter oxidative stress
levels in the dam and pup (3) however, maternal folic acid
supplementation in the absence of vitamin B12 increased
oxidative stress levels both in the dam and pup (4) mater-
nal folic acid supplementation both in the presence and
absence of vitamin B12 reduced brain DHA levels in the
pup (5) Maternal omega 3 fatty acids supplementation
to the vitamin B12 deficient groups resulted in signifi-
cantly higher brain DHA and lower AA levels.
In our study there was no difference in the total
weight gain of animals among all the groups. Our results
are similar to those by Achon et al. [38] who showed that
folic acid supplementation at 40 mg/kg diet did not
affect the gestation outcome. There are no reports which
have examined effects of folic acid supplementation in
68 S. Roy et al. / Brain & Development 34 (2012) 64–71

Fig. 1. Maternal plasma and pup brain malondialdehyde and fatty acid levels. *p < 0.05; **p < 0.01; compared to control; #p < 0.05; ##p < 0.01;
compared to NLFolate, noB12 $p < 0.05; $$p < 0.01; compared to HFolate, normal B12; @p < 0.05; @@p < 0.01; compared to HFolate,
9 99
noB12; xp < 0.05; xxp < 0.01 compared to NLFolate, noB12, omega 3 supp.; control – normal folate, normal B12; NLFolate, noB12 – normal
folate, B12 deficient; NLFolate, noB12, omega 3 supp – normal folate, B12 deficient, omega 3 supplemented HFolate, normal B12 – high folate, normal
B12; HFolate, noB12 – high folate, B12 deficient; HFolate, noB12, omega 3 supp – high folate, B12 deficient, omega 3 supplemented.

absence of vitamin B12 on birth outcome. However, in a was no difference in the body weights before and after
recent study when male rats were supplemented with supplementation [39]. In our study, omega 3 fatty acid
folic acid in the absence of vitamin B12 deficiency there did not influence the weight gain. Others also report that
 S. Roy et al. / Brain & Development 34 (2012) 64–71
varying ratios of omega 3 fatty:omega 6 fatty acids dur-
ing pregnancy on a control diet do not show any effect
on weight gain [40].
There was no difference in litter weight or size in all
groups due to altered levels of maternal micronutrients
in this study. Our earlier studies examining the effects
of folic acid supplementation (8 mg/kg) at marginal pro-
tein diet during pregnancy also found no change in litter
weight or litter size [41]. In contrast a study examining
the effect of supplementation of excess folic acid
(40 mg/kg) during pregnancy showed reductions in body
weight of the fetuses as compared to control [38]. In our
study when omega 3 fatty acid was given along with
excess folic acid in the absence of vitamin B12 there
was no change in pup weight. Maternal diets containing
varying ratios of omega 3 fatty:omega 6 fatty acids [40],
or docosahexaenoic acid and eicosapentaenoic acid [42]
have also shown no alterations in pup weights at birth.
However, when pregnant women were given 200 mg
DHA supplements from gestation week 21 until the
end of third month of lactation, their infants had lower
body weight [43]. In contrast maternal consumption of
omega 3 fatty acids during pregnancy have reported
an increase in birth weight [44–47].
It is known that the brain is vulnerable to free radical
attack and lipid peroxidation (LPO) [48]. There are no
studies which have examined the effect of imbalance of
maternal micronutrients or omega 3 fatty acid supple-
mentation to these diets on brain plasma MDA levels,
an index of lipid peroxidation. In our study maternal vita-
min B12 deficiency did not increase plasma MDA levels in
the dams. However, excess folic acid in the absence of
vitamin B12 increased plasma MDA levels. A recent study
in adult rats reports increased malondialdehyde (MDA)
levels in kidney tissue of vitamin B6 deficient male rats
[49] while folic acid and folic acid + vitamin B12 supple-
mentation in adult rats has also been shown to reduce sys-
temic oxidative stress in male rats [50]. Patients with
severe neonatal cobalamin (vitamin B12) deficiency have
been reported to have elevated intracellular ROS content
[51]. It has been suggested that dietary vitamin B12 folic
acid, and omega 3 PUFAs may influence plasma and
neuronal levels of oxidative stress, that may ultimately
determine the susceptibility of an individual to develop
neurodegenerative disorders [52].
It is known that LCPUFA is susceptible for
degradation due to increased oxidative stress [53].
The imbalances in micronutrients often lead to
increased oxidative stress through the generation of
oxygen free radicals which interact with polyunsatu-
rated fatty acids in membranes or lipoproteins.
Reactive oxygen species attack the double bonds of
polyunsaturated fatty acids and initiate chain reactions
leading to lipid peroxide formation. Our earlier studies
in women with pregnancy complications like pre-
eclampsia also suggest that increased peroxidative
69
reactions would further promote decomposition of
polyunsaturated fatty acids [25].
The present study for the first time reports lower pup
brain DHA levels in both the maternal vitamin B12 defi-
cient groups irrespective of the level of folic acid. We have
earlier reported that maternal folic acid supplementation
at marginal protein levels reduced brain DHA levels in
the adult offspring [20,54]. Altered intake/metabolism
of folic acid and vitamin B12 may induce oxidative stress
leading to increased MDA from LCPUFA.
Vitamin B12 and folate are essential for methylation of
homocysteine to methionine and for synthesis of s-aden-
osyl methionine. s-Adenosyl methionine is involved in
the metabolism of proteins, phospholipids and neuro-
transmitters [28]. This also can reduce levels of glutathi-
one, a major antioxidant during pregnancy in mother
and fetus. In the liver, phosphatidylcholine (PC) is also
synthesized by the sequential methylation of phosphati-
dylethanolamine (PE), which is catalyzed by phosphati-
dylethanolamine N-methyltransferase (PEMT) [55]. In
vitamin B deficiency, the amounts of methionine
available for phosphatidylcholine synthesis is reduced
and a lower methylation of phosphatidylethanolamine
to phosphatidylcholine has been reported [56]. A defect
in methylation process is hypothesized to be the biochem-
ical basis of neuropsychiatric manifestations of vitamin
B12 deficiency [57].
Folic acid and vitamin B12 are the major determi-
nants of one carbon metabolism which alter levels of
both homocysteine and oxidative stress (MDA). One
of the major functions of one carbon metabolism is also
to regulate the membrane phospholipids levels especially
DHA through the transfer of methyl groups. Therefore
altered intake/metabolism of maternal micronutrients
may alter levels of DHA (Fig. 2).
Supplementing the vitamin B12 deficient groups with
omega 3 fatty acids in our study resulted in higher levels
of brain DHA levels but reduced AA levels. A recent study
Fig. 2. Altered maternal micronutrient interactions leading to impaired
brain development in the offspring.
70 S. Roy et al. / Brain & Development 34 (2012) 64–71
has shown that early supplementation of maternal diet
with omega 3 fatty acids supplied as EPA and DHA is
efficient in increasing omega 3 in brain of the neonate
[42]. Several neurodevelopmental disorders like schizo-
phrenia [58] and ADHD [59] are associated with increased
oxidative stress. However there is no information when
and how oxidative stress triggered by an etiopathogenetic
factor(s) affects the early neurodevelopment and contrib-
utes to onset of psychopathology and its course and treat-
ment throughout life.
Our results suggest that oxidative stress is most likely
triggered due to imbalance in maternal micronutrients
like folic acid and vitamin B12 during fetal growth. This
may continue through adolescence thereby contributing
to neurodevelopmental deficits and onset of illness.
These micronutrients are involved in 1-carbon metabo-
lism and act as co-factors or molecular donors for
epigenetic processes such as DNA methylation, which
modulate gene expression, and therefore cellular prolif-
eration and apoptosis, in early pregnancy [3]. There is a
need to determine the underlying mechanisms of action
of these nutrients, including effects mediated via epige-
netic modifications. Our lab has therefore undertaken
studies to examine the epigenetic changes occurring at
the gene specific level for antioxidant enzymes (SOD
and GPx) and neurotrophins like brain derived neuro-
trophic factor which is known to be regulated by levels
of DHA [60,61] and these findings may have important
implications in neurodevelopmental disorders.
This study for the first time indicates that imbalance
in maternal micronutrients reduces the brain DHA lev-
els in the offspring at birth probably through the
observed increase in oxidative stress. Supplementation
with omega 3 fatty acids to these imbalanced diets
improved the brain DHA levels. Our data has implica-
tions for several neurodevelopmental disorders since
micronutrients and DHA are important modulators
for neural functioning. Therefore it would be of inter-
est to examine whether the observed alterations in
maternal micronutrients and the consequential reduc-
tions in brain fatty acids in the offspring’s are due to
the altered methylation and expression of PEMT.
Future studies need to also address the associated
changes in brain structure and function during the
adult life.
Conflict of interest statement
None.
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